Renal ischemia-reperfusion injury (IRI) is involved in the majority of clinical conditions that manifest as renal function deterioration; however, specific treatment for this type of injury is still far from clinical use. Since Toll-like receptor (TLR)-mediated signaling is a key mediator of IRI, we examined the effect of a multiple-TLR-blocking peptide named TLR-inhibitory peptide 1 (TIP1), which exerts the strongest action on TLR4, on renal IRI. We subjected C57BL/6 mice to 23 min of renal pedicle clamping preceded by intraperitoneal injection with a vehicle or TIP1. Sham control mice underwent flank incision only. Mouse kidneys were harvested after 24 h of reperfusion for histology, western blot, RT-PCR, and flow cytometry analysis. Pretreatment with TIP1 lowered the magnitude of elevated plasma creatinine levels and attenuated tubular injury. TIP1 treatment also reduced mRNA expression of inflammatory cytokines and decreased apoptotic cells and oxidative stress in post-ischemic kidneys. In kidneys pretreated with TIP1, the infiltration of macrophages and T helper 17 cells was less abundant than those in the IRI only group. These results suggest that TIP1 has a potential beneficial effect in attenuating the degree of kidney damage induced by IRI.

The diagnosis and post-treatment monitoring of head and neck squamous cell carcinoma (HNSCC) are challenging, highlighting the need for reliable and noninvasive biomarkers. Salivary exosomal microRNAs are promising candidates; however, their clinical utility has been hampered by limitations of conventional isolation methods. To address this gap, we employed a rigorous discovery-validation study design using independent patient cohorts, transitioning from ultracentrifugation in the discovery phase to an advanced, gentle hydrogel-based method for exosome enrichment in the validation phase. Candidate microRNAs were screened using microarray analysis and validated using quantitative real-time polymerase chain reaction. Our multistep screening identified five robustly detectable microRNAs. Among these, salivary exosomal miR-26a-5p has demonstrated a high accuracy in diagnosing HNSCC, particularly oral cavity cancers. Furthermore, post-operative decreases in miR-1290 and miR-4454 levels were strongly associated with favorable clinical outcomes, whereas persistently high or increased levels indicated a high risk of recurrence or poor prognosis. This study validated a panel of salivary exosomal microRNAs as powerful, non-invasive biomarkers, and the use of a novel hydrogel-based enrichment technique enhanced the reliability and clinical applicability of these findings to improve patient-management strategies.